RE: [Histonet] Staining coral tissues embedded in resin

From:

"Patsy Ruegg"

Laura,
For H&E's on GMA embedded tissue, you stain right thru the plastic, it is
water permeable and cannot be removed, there fore it will often take a more
concentrated and or longer incubation times to stain. I use Gill's III
hematoxylin for GMA sections for 10 min., you can use alcoholic eosin but
beware that alcohol with any water in it tends to lift the GMA section off
the slide. Here is what I do:
Gill's #3 10 min.
Tap wash for 1 min.
Dip in ammonia water to blue (10 dips)
Tap wash for 5 min.
I then airdry the sections before going to alcoholic eosin
Alcoholic eosin (right now I am using SurgiPath) 60 dips then quickly 5 dips
in 95% alcohol, then to 100% and xylene (once past the alcohol with water
the sections are fine. An alternative is to use aqueous eosin, rinse with
water, airdry and then dip one at a time in xylene and coverslip.
For all other stains that end in water rinses I airdry GMA sections and then
coverslip without going thru graded alcohols to dehydrate.
Hopes this helps,
Patsy
Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg@ihctech.net
web site www.ihctech.net
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-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laura Hunt
Sent: Wednesday, February 06, 2008 8:20 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Staining coral tissues embedded in resin
Hi everyone,
I am new to the forum. I am working on coral tissues embedded in
Immunobed resin and paraffin. We would like to stain both with H&E
and Azure II /Basic Fuchsin. I have never tried staining plastic with
H&E. Any suggestions or comments would be great! Since corals have a
calcium carbonate skeleton, the tissues were decalcified in a EDTA
solution.
Laura Hunt
UTA
On Feb 5, 2008, at 8:07 PM, histonet-request@lists.utsouthwestern.edu
wrote:
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>
> Today's Topics:
>
> 1. EpoR antibody (Colleen Forster)
> 2. RE: sections falling off (Debrosse-Serra, Beatrice)
> 3. Re: sections falling off (Rene J Buesa)
> 4. Re: Alcian Blue Quality Control OT (Robert Richmond)
> 5. HP staining (Martin, Gary)
> 6. Slide printers (ink) (Kim Merriam)
> 7. RE: Slide printers (ink) (Charles, Roger)
> 8. RE: Slide printers (ink) (Weems, Joyce)
> 9. Immunoperoxidase Double staining protocol (Reuel Cornelia)
> 10. Re: Slide printers (ink) (Patti Loykasek)
> 11. RE: Storage codes (Christine Tambasco)
> 12. Re: Slide printers (ink) (pkromund@gundluth.org)
> 13. Re: Immunoperoxidase Double staining protocol (Rene J Buesa)
> 14. thanks for the support and good wishes (Sebree Linda A.)
> 15. (no subject) (Margaryan, Naira)
> 16. Do mice have tonsils?? (Colleen Forster)
> 17. eosinophils on frozen section (Breisch, Eric)
> 18. RE: eosinophils on frozen section (Ingles Claire)
> 19. A gentle reminder about using subject line Re: [Histonet] (no
> subject) (Gayle Callis)
> 20. RE: eosinophils on frozen section (Weems, Joyce)
> 21. Re: Do mice have tonsils?? (Gayle Callis)
> 22. Re: eosinophils on frozen section (Gayle Callis)
> 23. RE: eosinophils on frozen section (Julia Dahl)
>
>
> ----------------------------------------------------------------------
>
> Message: 1
> Date: Tue, 05 Feb 2008 12:20:37 -0600
> From: Colleen Forster
> Subject: [Histonet] EpoR antibody
> To: histonet@lists.utsouthwestern.edu
> Message-ID: <47A8A8F5.3090606@umn.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
>
> Hello histonetters,
>
> Is anyone doing the EpoR antibody on FFPE for IHC? If so, could you
> share the vendor and a starting dilution.....thanks.
>
> Colleen Forster
> U of MN
>
>
>
> ------------------------------
>
> Message: 2
> Date: Tue, 5 Feb 2008 10:30:10 -0800
> From: "Debrosse-Serra, Beatrice"
> Subject: RE: [Histonet] sections falling off
> To: "Olek Michalski" , "Histonet"
>
> Message-ID:
>
> <8404DFBED5207B4B8E5EEF4332CEEA5305BD4C99@lajamrexm01.amer.pfizer.com>
> Content-Type: text/plain; charset="us-ascii"
>
> Try Newcomers silane coated slides. They are not cheap, but the
> sections
> stay on very well.
>
> Beatrice DeBrosse-Serra
> Pathology Scientist
> Pfizer Global Research & Development
> CB4, 2150
> 10646 Science Center Drive
> San Diego, CA 92121
> Phone# 858-622-5986
> Fax# 858-678-8290
>
>
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Olek
> Michalski
> Sent: Tuesday, February 05, 2008 10:00 AM
> To: Histonet
> Subject: [Histonet] sections falling off
>
>
> Dear Histonetters,
>
> a friend of mine just faced a massive sections falling off. She is
> doing
>
> Nissl staining in mouse brain (brains perfused with formalin,
> postfixed
> 3
> days, and cryosectioned) on poly-L-lysined slides. She just managed to
> pass trough xylene and decreasing grades of alcohol to water and the
> sections started to detach. We were trying to reattach them but it
> went
>
> out that sections are not flat any more. It seems like the tissue is
> rehydrated unevenly, but keeping it in water for hours didn't work. Is
> there any way to spread these sections not damaging them at the same
> time?
> She is likely to rescue this sections even if some work is needed.
>
> Best regards
> Olek Michalski
> --
> Laboratory of Neurobiology
> of Development and Evolution
>
> Nencki Institute of Experimental Biology
> ul. Pasteura 3, 02-093 Warszawa, Poland
> Tel. +48 22 5892268, Fax +48 22 8225342
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 3
> Date: Tue, 5 Feb 2008 10:30:54 -0800 (PST)
> From: Rene J Buesa
> Subject: Re: [Histonet] sections falling off
> To: Olek Michalski , Histonet
>
> Message-ID: <87465.99172.qm@web61214.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> If they were cryosectioned she did not have to go through xylene or
> graded alcohols to "dewax and hydrate" since there was no
> dehydration or wax infiltration to begin with.
>
> Cryosectioned sections just need to wash away the medium used to
> cryosection (OCT perhaps?) and just stain with the aqueous solutions.
>
> Later they can de dehydrated and cleared if a permanent mount is
> desired.
> Try to float the sections in a water bath and pick them up and
> don't try again to use xylene or alcohols.
> Probably she was following a procedure for paraffin embedded tissue
> that should have been adapted to frozen sections and it was not.
> René J.
>
> Olek Michalski wrote:
>
> Dear Histonetters,
>
> a friend of mine just faced a massive sections falling off. She is
> doing
> Nissl staining in mouse brain (brains perfused with formalin,
> postfixed 3
> days, and cryosectioned) on poly-L-lysined slides. She just managed to
> pass trough xylene and decreasing grades of alcohol to water and the
> sections started to detach. We were trying to reattach them but it
> went
> out that sections are not flat any more. It seems like the tissue is
> rehydrated unevenly, but keeping it in water for hours didn't work. Is
> there any way to spread these sections not damaging them at the same
> time?
> She is likely to rescue this sections even if some work is needed.
>
> Best regards
> Olek Michalski
> --
> Laboratory of Neurobiology
> of Development and Evolution
>
> Nencki Institute of Experimental Biology
> ul. Pasteura 3, 02-093 Warszawa, Poland
> Tel. +48 22 5892268, Fax +48 22 8225342
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ---------------------------------
> Looking for last minute shopping deals? Find them fast with Yahoo!
> Search.
>
> ------------------------------
>
> Message: 4
> Date: Tue, 5 Feb 2008 13:45:34 -0500
> From: "Robert Richmond"
> Subject: [Histonet] Re: Alcian Blue Quality Control OT
> To: histonet@lists.utsouthwestern.edu
> Message-ID:
>
> Content-Type: text/plain; charset=ISO-8859-1
>
> Claire Ingles remarks: So that's what GOP means!
>
> Not this grumpy old pathologist - I voted for Barack Obama!
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
>
>
>
> ------------------------------
>
> Message: 5
> Date: Tue, 5 Feb 2008 11:00:57 -0800
> From: "Martin, Gary"
> Subject: [Histonet] HP staining
> To:
> Message-ID:
> <6ED9D4252F278841A0593D3D788AF24C01ACDF72@mailsvr.MARSHMED.local>
> Content-Type: text/plain; charset="us-ascii"
>
> To all who responded to my e-mail concerning rapid HP staining to
> include a yellow back ground ... thank you very much ... my question
> was
> answered.
>
> Gray
>
>
>
> ------------------------------
>
> Message: 6
> Date: Tue, 5 Feb 2008 11:48:08 -0800 (PST)
> From: Kim Merriam
> Subject: [Histonet] Slide printers (ink)
> To: Histonet
> Message-ID: <813260.30617.qm@web50306.mail.re2.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Hi everyone,
>
> I was wondering what everyone's experience was with the TBS slide
> printer (Shurmark-plus), the one that prints with ink? I would
> appreciate input, good or bad.
>
> We currently have an etcher, and we hate it, so we are looking to
> trade it in for something that uses ink. I have used the Leica
> printer in the past and have had reasonably good luck with it, but I
> was wondering about the printer from TBS, I don't know anyone that
> has one.
>
> Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
> Try it now.
>
> ------------------------------
>
> Message: 7
> Date: Tue, 5 Feb 2008 14:58:42 -0500
> From: "Charles, Roger"
> Subject: RE: [Histonet] Slide printers (ink)
> To: "Kim Merriam"
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID:
> <12E4E17FEF6EBE4BAE95BEB3CDCB57000943BB6E@enhbgpri04.backup>
> Content-Type: text/plain; charset="US-ASCII"
>
> Hello,
> Recently we bought a sakura slide writer that uses ink to print. The
> only problem I have found with the ink is if you put labels over the
> print and then try to remove the label to verify a number error, or
> something like that, the ink will actually come off with the label.
> If
> you can read upside down you can still verify the number. I did run
> this ink thru a battery of chemicals including straight formic acid
> for
> 5 minutes and the ink did not leave the slide. I would caution in
> buying a Sakura slide write as it has a very unfriendly Access based
> program and the writer does not work with charged slides. This has
> been
> verified by me and an independent engineer sent to look at this issue.
> I do use a TBS cassette writer and love that so if I can get my sakura
> sent back I will be looking at TBS's slide writer too.
> Good luck
> Roger
>
> Roger Charles
> Microbiologist
> Pennsylvania Veterinary Laboratory
> 2305 N Cameron St
> Harrisburg, PA 17110
> 717-787-8808
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim
> Merriam
> Sent: Tuesday, February 05, 2008 2:48 PM
> To: Histonet
> Subject: [Histonet] Slide printers (ink)
>
> Hi everyone,
>
> I was wondering what everyone's experience was with the TBS slide
> printer (Shurmark-plus), the one that prints with ink? I would
> appreciate input, good or bad.
>
> We currently have an etcher, and we hate it, so we are looking to
> trade it in for something that uses ink. I have used the Leica
> printer
> in the past and have had reasonably good luck with it, but I was
> wondering about the printer from TBS, I don't know anyone that has
> one.
>
> Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
> Try
> it now.
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
> ------------------------------
>
> Message: 8
> Date: Tue, 5 Feb 2008 15:25:58 -0500
> From: "Weems, Joyce"
> Subject: RE: [Histonet] Slide printers (ink)
> To: "Charles, Roger" , "Kim Merriam"
>
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID:
> <1CD6831EB9B26D45B0A3EAA79F7EBD320518E54D@sjhaexc02.sjha.org>
> Content-Type: text/plain; charset="us-ascii"
>
> We have Sakura cassette and slide writers. We print charged slides
> daily.
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of
> Charles,
> Roger
> Sent: Tuesday, February 05, 2008 2:59 PM
> To: Kim Merriam
> Cc: histonet@lists.utsouthwestern.edu
> Subject: RE: [Histonet] Slide printers (ink)
>
> Hello,
> Recently we bought a sakura slide writer that uses ink to print. The
> only problem I have found with the ink is if you put labels over the
> print and then try to remove the label to verify a number error, or
> something like that, the ink will actually come off with the label.
> If
> you can read upside down you can still verify the number. I did run
> this ink thru a battery of chemicals including straight formic acid
> for
> 5 minutes and the ink did not leave the slide. I would caution in
> buying a Sakura slide write as it has a very unfriendly Access based
> program and the writer does not work with charged slides. This has
> been
> verified by me and an independent engineer sent to look at this issue.
> I do use a TBS cassette writer and love that so if I can get my sakura
> sent back I will be looking at TBS's slide writer too.
> Good luck
> Roger
>
> Roger Charles
> Microbiologist
> Pennsylvania Veterinary Laboratory
> 2305 N Cameron St
> Harrisburg, PA 17110
> 717-787-8808
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim
> Merriam
> Sent: Tuesday, February 05, 2008 2:48 PM
> To: Histonet
> Subject: [Histonet] Slide printers (ink)
>
> Hi everyone,
>
> I was wondering what everyone's experience was with the TBS slide
> printer (Shurmark-plus), the one that prints with ink? I would
> appreciate input, good or bad.
>
> We currently have an etcher, and we hate it, so we are looking to
> trade it in for something that uses ink. I have used the Leica
> printer
> in the past and have had reasonably good luck with it, but I was
> wondering about the printer from TBS, I don't know anyone that has
> one.
>
> Kim
>
>
> Kim Merriam, MA, HT(ASCP)
> Cambridge, MA
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
> Try
> it now.
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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>
>
> ------------------------------
>
> Message: 9
> Date: Tue, 05 Feb 2008 14:34:27 -0600
> From: "Reuel Cornelia"
> Subject: [Histonet] Immunoperoxidase Double staining protocol
> To:
> Message-ID: <47A873F3020000C50002ADF9@nwcl02.tsrh.org>
> Content-Type: text/plain; charset=US-ASCII
>
> I was asked to do an immunoperoxidase double staining on CD61 ,Cd42,
> pf4 with an unknown mouse antibody that will work on platelets. Can
> somebody help me or any protocol available that I can refer to. I am
> used of IF double stianing but not Immunoperoxidase. Thank you.
>
> Reuel Cornelia, BS MT, AMT
> Cellular Pathology
> Texas Scottish Rite Hospital for Children
> 2222 Welborn Street
> Dallas, TX 75219
> Tel: 214-559-7766
> fax: 214-559-7768
>
>
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>
> ------------------------------
>
> Message: 10
> Date: Tue, 05 Feb 2008 12:39:50 -0800
> From: Patti Loykasek
> Subject: Re: [Histonet] Slide printers (ink)
> To: "Charles, Roger" , Kim Merriam
>
> Cc: histonet@lists.utsouthwestern.edu
> Message-ID:
> Content-Type: text/plain; charset="US-ASCII"
>
> Roger- We have been using a sakura slide printer for a couple of
> years. I do
> agree with your comments about the software - not user friendly at
> all nor
> is it intuitive. I have used Access database in the past without these
> issues. I do not know about the labels, I don't think we've had that
> issue.
> Maybe it varies with the label. I don't quite understand your
> comment on
> charged slides, as all of the slides we put thru the printer are
> charged and
> we have no issues with that. We use charged slides (Okando Plus)
> from BBC
> and Probe-On Plus slides from ThermoFisher. What brand of slides
> were you
> using & what issue did you have with the slide printer from the usage?
>
>
> Patti Loykasek BS, HTL, QIHC
> PhenoPath Laboratories
> Seattle, WA
>
>
>
>
>
>
>
>> Hello,
>> Recently we bought a sakura slide writer that uses ink to print. The
>> only problem I have found with the ink is if you put labels over the
>> print and then try to remove the label to verify a number error, or
>> something like that, the ink will actually come off with the
>> label. If
>> you can read upside down you can still verify the number. I did run
>> this ink thru a battery of chemicals including straight formic acid
>> for
>> 5 minutes and the ink did not leave the slide. I would caution in
>> buying a Sakura slide write as it has a very unfriendly Access based
>> program and the writer does not work with charged slides. This has
>> been
>> verified by me and an independent engineer sent to look at this
>> issue.
>> I do use a TBS cassette writer and love that so if I can get my
>> sakura
>> sent back I will be looking at TBS's slide writer too.
>> Good luck
>> Roger
>>
>> Roger Charles
>> Microbiologist
>> Pennsylvania Veterinary Laboratory
>> 2305 N Cameron St
>> Harrisburg, PA 17110
>> 717-787-8808
>>
>> -----Original Message-----
>> From: histonet-bounces@lists.utsouthwestern.edu
>> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim
>> Merriam
>> Sent: Tuesday, February 05, 2008 2:48 PM
>> To: Histonet
>> Subject: [Histonet] Slide printers (ink)
>>
>> Hi everyone,
>>
>> I was wondering what everyone's experience was with the TBS slide
>> printer (Shurmark-plus), the one that prints with ink? I would
>> appreciate input, good or bad.
>>
>> We currently have an etcher, and we hate it, so we are looking to
>> trade it in for something that uses ink. I have used the Leica
>> printer
>> in the past and have had reasonably good luck with it, but I was
>> wondering about the printer from TBS, I don't know anyone that has
>> one.
>>
>> Kim
>>
>>
>> Kim Merriam, MA, HT(ASCP)
>> Cambridge, MA
>>
>> ---------------------------------
>> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
>> Try
>> it now.
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
>
>
> This e-mail message, including any attachments, is for the sole use
> of the
> intended recipients and may contain privileged information. Any
> unauthorized
> review, use, disclosure or distribution is prohibited. If you are
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> recipient, please contact the sender by e-mail and destroy all
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> at (206) 374-9000.
>
>
>
>
> ------------------------------
>
> Message: 11
> Date: Tue, 5 Feb 2008 21:10:37 +0000
> From: Christine Tambasco
> Subject: RE: [Histonet] Storage codes
> To: Barbara Albert , histonet
>
> Message-ID:
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> I always kept my gold chloride in the refrigerator. Yours is in a
> vial though so is it a powder?
> That is what I used to have I kept at room temperature.
> Just my advice.
> Christine Tambasco, HT (ASCP)
> St. Marys Hospital
> Amsterdam, NY> Date: Mon, 4 Feb 2008 15:33:30 -0800> From:
barbaraaalbert@yahoo.com
> > To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] Storage
> codes> > Hi all,> I just received some vials of Gold Chloride that I
> had ordered and was checking the label for stoarge requirements. It
> has "Storage Code White". I haven't heard of storage codes and want
> to know where I can go to find out what they mean.> > Thanks,>
> Barbara Albert> UCSF Medical Center> San Francisco> > >
> ---------------------------------> Never miss a thing. Make Yahoo
> your homepage.> _______________________________________________>
> Histonet mailing list> Histonet@lists.utsouthwestern.edu>
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
> _________________________________________________________________
> Climb to the top of the charts! Play the word scramble challenge
> with star power.
> http://club.live.com/star_shuffle.aspx?icid=starshuffle_wlmailtextlink_jan
>
> ------------------------------
>
> Message: 12
> Date: Tue, 5 Feb 2008 15:34:42 -0600
> From: pkromund@gundluth.org
> Subject: Re: [Histonet] Slide printers (ink)
> To: Patti Loykasek
> Cc: histonet@lists.utsouthwestern.edu,
> histonet-bounces@lists.utsouthwestern.edu, "Charles, Roger"
>
> Message-ID:
>
>
>
> Content-Type: text/plain; charset=US-ASCII
>
> Do any of these slide printers affect precut control tissue on the
> charged
> slides?
> Pamela
>
>
>
> Patti Loykasek
>
> ath.com> To
> Sent by: "Charles, Roger"
> histonet-bounces@ , Kim
> Merriam
> lists.utsouthwest
>
> ern.edu cc
>
> histonet@lists.utsouthwestern.edu
>
> Subject
> 02/05/2008 02:39 Re: [Histonet] Slide printers
> (ink)
> PM
>
>
>
>
>
>
>
>
>
> Roger- We have been using a sakura slide printer for a couple of
> years. I
> do
> agree with your comments about the software - not user friendly at
> all nor
> is it intuitive. I have used Access database in the past without these
> issues. I do not know about the labels, I don't think we've had that
> issue.
> Maybe it varies with the label. I don't quite understand your
> comment on
> charged slides, as all of the slides we put thru the printer are
> charged
> and
> we have no issues with that. We use charged slides (Okando Plus)
> from BBC
> and Probe-On Plus slides from ThermoFisher. What brand of slides
> were you
> using & what issue did you have with the slide printer from the usage?
>
>
> Patti Loykasek BS, HTL, QIHC
> PhenoPath Laboratories
> Seattle, WA
>
>
>
>
>
>
>
>> Hello,
>> Recently we bought a sakura slide writer that uses ink to print. The
>> only problem I have found with the ink is if you put labels over the
>> print and then try to remove the label to verify a number error, or
>> something like that, the ink will actually come off with the
>> label. If
>> you can read upside down you can still verify the number. I did run
>> this ink thru a battery of chemicals including straight formic acid
>> for
>> 5 minutes and the ink did not leave the slide. I would caution in
>> buying a Sakura slide write as it has a very unfriendly Access based
>> program and the writer does not work with charged slides. This has
>> been
>> verified by me and an independent engineer sent to look at this
>> issue.
>> I do use a TBS cassette writer and love that so if I can get my
>> sakura
>> sent back I will be looking at TBS's slide writer too.
>> Good luck
>> Roger
>>
>> Roger Charles
>> Microbiologist
>> Pennsylvania Veterinary Laboratory
>> 2305 N Cameron St
>> Harrisburg, PA 17110
>> 717-787-8808
>>
>> -----Original Message-----
>> From: histonet-bounces@lists.utsouthwestern.edu
>> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim
>> Merriam
>> Sent: Tuesday, February 05, 2008 2:48 PM
>> To: Histonet
>> Subject: [Histonet] Slide printers (ink)
>>
>> Hi everyone,
>>
>> I was wondering what everyone's experience was with the TBS slide
>> printer (Shurmark-plus), the one that prints with ink? I would
>> appreciate input, good or bad.
>>
>> We currently have an etcher, and we hate it, so we are looking to
>> trade it in for something that uses ink. I have used the Leica
>> printer
>> in the past and have had reasonably good luck with it, but I was
>> wondering about the printer from TBS, I don't know anyone that has
>> one.
>>
>> Kim
>>
>>
>> Kim Merriam, MA, HT(ASCP)
>> Cambridge, MA
>>
>> ---------------------------------
>> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
>> Try
>> it now.
>> _______________________________________________
>> Histonet mailing list
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>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
>
>
> This e-mail message, including any attachments, is for the sole use
> of the
> intended recipients and may contain privileged information. Any
> unauthorized
> review, use, disclosure or distribution is prohibited. If you are
> not the
> intended
> recipient, please contact the sender by e-mail and destroy all
> copies of
> the
> original message, or you may call PhenoPath Laboratories, Seattle, WA
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> at (206) 374-9000.
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>
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>
>
>
>
> ------------------------------
>
> Message: 13
> Date: Tue, 5 Feb 2008 13:46:41 -0800 (PST)
> From: Rene J Buesa
> Subject: Re: [Histonet] Immunoperoxidase Double staining protocol
> To: Reuel Cornelia ,
> histonet@lists.utsouthwestern.edu
> Message-ID: <784337.61146.qm@web61225.mail.yahoo.com>
> Content-Type: text/plain; charset=iso-8859-1
>
> Generally speaking you will have to run the whole protocol for one
> of the Abs and use one chromogen (of your selection). After that,
> since you already did HIER and blocked the internal peroxidase, you
> will treat the section with the second Ab, will link/detect it and
> use another chromogen that ideally should have a different color.
> The first chromogen could be regular DAB and for the second you can
> use also DAB but with incorporated nickel that will give a deep/blue
> purple color, in contrast with the brown/reddish normal color of the
> DAB.
> The thing is that you don't need any special protocol, just your
> regular IHC protocol for FFPE tissue, run twice with two different
> chromogens.
> René J.
>
> Reuel Cornelia wrote:
> I was asked to do an immunoperoxidase double staining on
> CD61 ,Cd42, pf4 with an unknown mouse antibody that will work on
> platelets. Can somebody help me or any protocol available that I can
> refer to. I am used of IF double stianing but not Immunoperoxidase.
> Thank you.
>
> Reuel Cornelia, BS MT, AMT
> Cellular Pathology
> Texas Scottish Rite Hospital for Children
> 2222 Welborn Street
> Dallas, TX 75219
> Tel: 214-559-7766
> fax: 214-559-7768
>
>
****************************************************************************
***************************************
> Texas Scottish Rite Hospital for Children is one of the nation's
> leading pediatric centers for the
> treatment of orthopedic conditions, certain related neurological
> disorders and learning disorders,
> such as dyslexia. This email transmission and/or its attachments may
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> information, intended only for the use of the individual or entity
> named above.
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> The authorized recipient of this information is prohibited from
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> _______________________________________________
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> Histonet@lists.utsouthwestern.edu
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>
>
>
> ---------------------------------
> Be a better friend, newshound, and know-it-all with Yahoo! Mobile.
> Try it now.
>
> ------------------------------
>
> Message: 14
> Date: Tue, 5 Feb 2008 15:47:03 -0600
> From: "Sebree Linda A."
> Subject: [Histonet] thanks for the support and good wishes
> To: "Histonet"
> Message-ID:
>
>
>
> Content-Type: text/plain; charset="US-ASCII"
>
> Hi all,
>
> Well apparently our CAP inspector took the word of our supervisor on
> everything pertaining to our lab, reviewed some of our slides and was
> out the door before the weather associated with the "winter storm
> warning" arrived! Some times its good to live in the land of ice and
> snow!
>
> Thanks for all the good luck wishes!
>
> Linda Sebree, HT(ASCP)
> University of Wisconsin Hospital & Clinics
> IHC/ISH Laboratory
> A4/204-3224
> 600 Highland Ave.
> Madison, WI 53792
> (608)265-6596
> FAX: (608)262-7174
>
>
>
> ------------------------------
>
> Message: 15
> Date: Tue, 5 Feb 2008 16:50:57 -0600
> From: "Margaryan, Naira"
> Subject: [Histonet] (no subject)
> To:
> Message-ID:
>
>
>
> Content-Type: text/plain; charset="us-ascii"
>
> Dear histo team,
>
>
>
> I need to stain frozen & FFPE sections for Tuj-1 (AbCam, ab53234) and
> FFPE sectiond for Netrin-1 (AbCam, ab39370). Any kind of suggestions
> or
> even full IHC or IF protocol is appreciated.
>
>
>
> Thanks in advance,
>
> Naira
>
>
>
>
>
>
>
>
>
> ------------------------------
>
> Message: 16
> Date: Tue, 05 Feb 2008 17:36:48 -0600
> From: Colleen Forster
> Subject: [Histonet] Do mice have tonsils??
> To: histonet@lists.utsouthwestern.edu
> Message-ID: <47A8F310.20905@umn.edu>
> Content-Type: text/plain; charset=ISO-8859-1; format=flowed
>
> For any of you have done mouse necropsy, do they have tonsils or just
> lymph nodes? I have had a request for mouse tonsils.... can't seem to
> find them in my searches. I would appreciate any help on this one.
>
> Thanks,
>
> Colleen Forster
> U of MN
>
>
>
> ------------------------------
>
> Message: 17
> Date: Tue, 5 Feb 2008 15:32:31 -0800
> From: "Breisch, Eric"
> Subject: [Histonet] eosinophils on frozen section
> To:
> Message-ID:
> <43B97B4C402C2C44AAA2A8D2C86A88B31A5A78@e2k3backend1.RCHSD.org>
> Content-Type: text/plain; charset="US-ASCII"
>
> To all interested Histonetters:
>
>
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success
> identifying
> eosinophils from a frozen section? We have had many frozen sections
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,
> three
> rinses in Xylene substitute and then coverslip. All other tissues
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
>
>
> ------------------------------
>
> Message: 18
> Date: Tue, 5 Feb 2008 18:08:15 -0600
> From: "Ingles Claire"
> Subject: RE: [Histonet] eosinophils on frozen section
> To:
> Message-ID:
>
<08A0A863637F1349BBFD83A96B27A50A1200DF@uwhis-xchng3.uwhis.hosp.wisc.edu
> >
>
> Content-Type: text/plain; charset="iso-8859-1"
>
> Eric:
> My guess is that the alcohol is lysing(blowing up) the unfixed
> eosinophils, therefore they are not there to stain. Fixation
> stabilizes the cells in order to be stained in regular paraffin
> sections. Unfortunately, I have done a few trials using different
> fixatives for fresh tissue to see if I could preserve these little
> suckers. Nothing really worked satisfactorally. You might try fixing
> in formalin for a minute or two before you continue on with your
> stain and see if this helps. (don't forget to wash the sections
> afterward) Hope this helps.
> Claire
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success
> identifying
> eosinophils from a frozen section? We have had many frozen sections
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,
> three
> rinses in Xylene substitute and then coverslip. All other tissues
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 19
> Date: Tue, 5 Feb 2008 17:30:17 -0700
> From: "Gayle Callis"
> Subject: A gentle reminder about using subject line Re: [Histonet] (no
> subject)
> To: "Margaryan, Naira" ,
>
> Message-ID: <000601c86857$72c69700$6601a8c0@Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> reply-type=original
>
> For those new to or not used to the Histonet messaging, please use the
> subject line rather ant just hit the reply key. Replying to the daily
> digest also kicks back all the messages for that day to those who have
> already received them. The subject is important so others, including
> me,
> don't just hit the delete key and not read your valuable inquiries and
> comment.
>
> Thanks
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
> ----- Original Message -----
> From: "Margaryan, Naira"
> To:
> Sent: Tuesday, February 05, 2008 3:50 PM
> Subject: [Histonet] (no subject)
>
>
> Dear histo team,
>
>
>
> I need to stain frozen & FFPE sections for Tuj-1 (AbCam, ab53234) and
> FFPE sectiond for Netrin-1 (AbCam, ab39370). Any kind of suggestions
> or
> even full IHC or IF protocol is appreciated.
>
>
>
> Thanks in advance,
>
> Naira
>
>
>
>
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 20
> Date: Tue, 5 Feb 2008 19:35:12 -0500
> From: "Weems, Joyce"
> Subject: RE: [Histonet] eosinophils on frozen section
> To: "Ingles Claire" ,
>
> Message-ID:
> <1CD6831EB9B26D45B0A3EAA79F7EBD320518E56A@sjhaexc02.sjha.org>
> Content-Type: text/plain; charset="us-ascii"
>
> Fix in formal-alcohol (no, it is not wearing a tuxedo) 90 ml absolute
> alcohol - 10 ml 37% formaldehyde... Should work ok then... j
>
> -----Original Message-----
> From: histonet-bounces@lists.utsouthwestern.edu
> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles
> Claire
> Sent: Tuesday, February 05, 2008 7:08 PM
> To: histonet@lists.utsouthwestern.edu
> Subject: RE: [Histonet] eosinophils on frozen section
>
> Eric:
> My guess is that the alcohol is lysing(blowing up) the unfixed
> eosinophils, therefore they are not there to stain. Fixation
> stabilizes
> the cells in order to be stained in regular paraffin sections.
> Unfortunately, I have done a few trials using different fixatives for
> fresh tissue to see if I could preserve these little suckers. Nothing
> really worked satisfactorally. You might try fixing in formalin for a
> minute or two before you continue on with your stain and see if this
> helps. (don't forget to wash the sections afterward) Hope this helps.
> Claire
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success
> identifying
> eosinophils from a frozen section? We have had many frozen sections
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,
> three
> rinses in Xylene substitute and then coverslip. All other tissues
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
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>
>
> ------------------------------
>
> Message: 21
> Date: Tue, 5 Feb 2008 17:49:57 -0700
> From: "Gayle Callis"
> Subject: Re: [Histonet] Do mice have tonsils??
> To: "Colleen Forster" ,
>
> Message-ID: <000e01c8685a$321bd7d0$6601a8c0@Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> reply-type=response
>
> Colleen,
>
> Rodents do NOT have tonsils, but have tissues analogous (Sp?) to
> tonsils,
> called nasal associated lymphoid tissues located in a rather
> difficult spot
> at the back of the nasal turbinates. I suggest you get into the
> literature
> and type in murine NALT as there are some excellent articles on
> morphology
> and their location. If you want to do frozen sections, you either
> need to
> remove these from this area, not easy to do, and takes a lot of
> practice OR
> you can do undecalcified bone frozen sections with the Cryojane.
> The NALT
> is located above the soft palate, just below the 3rd palatine ridge
> if you
> start counting ridges on the soft palate and starting count at the
> front
> incisors. These tiny lymphoid tissues are nestled on the roof of
> the mouth,
> just below the soft palate, between the back molars. The molars are
> the
> complicating factor for doing murine CD markers that only work on
> frozen
> sections. I will be happy to send you a powerpoint photograph of a
> fully
> decalcified mouse head, mid sagittal section, stained with H&E to show
> exactly where the NALT is located. I also have a cross section of the
> murine nasal turbinates showing NALT in that orientation, and much
> harder to
> deal with in order to find the NALT. One project had PLP perfused
> mouse
> head, immersion fixed longer and then EDTA decalcified,
> cryoprotected and
> sectioned on the cryostat to do immunoglobulin staining of turbinate
> epithelial cells back and into the NALT.
>
> We work with NALT a good deal, and I can honestly say that
> dissection/removal is not easy. However, it can be done but with very
> gentle, light touch using a pointed, tiny scalpel blade. When we do
> frozen
> sections of removed NALT, we can obtain approx 30 - 40 sections,
> serial, at
> 5 um and with treated animals to produce inflammed NALT, up to 50
> and more
> serial sections. With undecalcified bone, we get far fewer due to the
> difficulty of sectioning.
>
> G0 into PUBMED, and look for David Pascual, Keri Cscentis on their
> NALT
> publication. I believe they put a cartoon of NALT location in that
> publication.
>
> Good luck,
>
> Gayle M. Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
> ----- Original Message -----
> From: "Colleen Forster"
> To:
> Sent: Tuesday, February 05, 2008 4:36 PM
> Subject: [Histonet] Do mice have tonsils??
>
>
>> For any of you have done mouse necropsy, do they have tonsils or just
>> lymph nodes? I have had a request for mouse tonsils.... can't seem
>> to find
>> them in my searches. I would appreciate any help on this one.
>>
>> Thanks,
>>
>> Colleen Forster
>> U of MN
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 22
> Date: Tue, 5 Feb 2008 17:58:05 -0700
> From: "Gayle Callis"
> Subject: Re: [Histonet] eosinophils on frozen section
> To: "Breisch, Eric" ,
>
> Message-ID: <001401c8685b$55212f90$6601a8c0@Sunney>
> Content-Type: text/plain; format=flowed; charset="iso-8859-1";
> reply-type=original
>
> We improved our eosinophil staining on frozen sections of murine
> tissue by
> switching to NBF immersion fixation, let it sit 10 min, then do the
> H&E but
> use eosin/phloxine (Richard Allan stain will work) but stain in
> eosin/phloxine for 2 minutes. For some reason, the eosin was
> never taken
> up as well after a shorter time in eosin alone, and greatly
> improved with
> the phloxine added. Sections can be cut a bit thinner too, try 4 um
> - it
> should make the granules more apparent against the background and
> other
> cells. Also, I suggest adding absolute alcohol after the 95% for
> better
> dehydration before clearing.
>
> You may have success using alcoholic formalin in order to speed up the
> fixation a bit. However, when we switched to NBF, we had better
> staining of
> eosinophils in frozen sections. If you have the time, you can let NBF
> fixation go longer, we often let sections sit for days then stain,
> but with
> your diagnostic procedure, you will want a bit more speed involved.
>
> Good luck
>
> Gayle M..Callis
> HTL/HT/MT(ASCP)
> Bozeman MT 59715
>
>
>
>
>
>
> ----- Original Message -----
> From: "Breisch, Eric"
> To:
> Sent: Tuesday, February 05, 2008 4:32 PM
> Subject: [Histonet] eosinophils on frozen section
>
>
> To all interested Histonetters:
>
>
>
>
>
> We are experiencing some frustration with our ability to identify
> eosinophils from frozen sections. For some reason which our frozen
> section set up does not enable us to identify eosinophils. Eosinophil
> identification is not impaired when the tissue is submitted for
> permanents. Could some one please share the reagents used and how the
> frozen section staining area is set up if they have success
> identifying
> eosinophils from a frozen section? We have had many frozen sections
> done
> on fresh tissues suspicious for an eosinophilic granuloma but our
> techniques so far have not rendered consistent eosinophil
> identification. We presently are using 95% ETOH for fixing the FS
> slide,
> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in 70%
> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,
> three
> rinses in Xylene substitute and then coverslip. All other tissues
> appear
> just fine but the eosinophils just don't show up. Any suggestions are
> greatly appreciated.
>
>
>
>
>
> Eric A. Breisch, Ph.D.
>
> Clinical Anatomist
>
> Dept. of Pathology
>
> Rady Children's Hospital and Health Center
>
> Associate Clinical Professor of Anatomy
>
> Dept. of Surgery
>
> UCSD School of Medicine
>
>
>
> _______________________________________________
> Histonet mailing list
> Histonet@lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>
>
> ------------------------------
>
> Message: 23
> Date: Tue, 5 Feb 2008 20:06:55 -0600
> From: Julia Dahl
> Subject: RE: [Histonet] eosinophils on frozen section
> To: "Breisch, Eric" ,
>
> Message-ID:
> Content-Type: text/plain; charset="iso-8859-1"
>
>
> Agree with Gayle that alcoholic formalin allows for speedier
> fixation and preservation of eosinophil granule membranes (and
> therefore staining, rather than degranulation).
>
>> Date: Tue, 5 Feb 2008 15:32:31 -0800
>> From: ebreisch@rchsd.org
>> To: histonet@lists.utsouthwestern.edu
>> Subject: [Histonet] eosinophils on frozen section
>>
>> To all interested Histonetters:
>>
>>
>>
>>
>>
>> We are experiencing some frustration with our ability to identify
>> eosinophils from frozen sections. For some reason which our frozen
>> section set up does not enable us to identify eosinophils. Eosinophil
>> identification is not impaired when the tissue is submitted for
>> permanents. Could some one please share the reagents used and how the
>> frozen section staining area is set up if they have success
>> identifying
>> eosinophils from a frozen section? We have had many frozen sections
>> done
>> on fresh tissues suspicious for an eosinophilic granuloma but our
>> techniques so far have not rendered consistent eosinophil
>> identification. We presently are using 95% ETOH for fixing the FS
>> slide,
>> quick wash in distilled H2O, 1 minute in Richard Allen Hematoxylin 2,
>> rinse in tap water, 15 seconds in bluing, rinse in water, rinse in
>> 70%
>> ETOH, 1 minute in Richard Allen Eosin Y, three rinses in 95% ETOH,
>> three
>> rinses in Xylene substitute and then coverslip. All other tissues
>> appear
>> just fine but the eosinophils just don't show up. Any suggestions are
>> greatly appreciated.
>>
>>
>>
>>
>>
>> Eric A. Breisch, Ph.D.
>>
>> Clinical Anatomist
>>
>> Dept. of Pathology
>>
>> Rady Children's Hospital and Health Center
>>
>> Associate Clinical Professor of Anatomy
>>
>> Dept. of Surgery
>>
>> UCSD School of Medicine
>>
>>
>>
>> _______________________________________________
>> Histonet mailing list
>> Histonet@lists.utsouthwestern.edu
>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
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> End of Histonet Digest, Vol 51, Issue 7
> ***************************************
Laura R Hunt, PhD
Post-doctoral associate
University of Texas at Arlington
ph: 817-272-1499
fax: 817-272-2855
lhunt@uta.edu
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